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Module 5

Module 5. Laboratory Diagnostics, Specimen Collection, and Biosafety Issues. Learning Objectives. Prepare and maintain collection kits Collect and transport specimens safely and correctly Manage laboratory specimens safely Interpret laboratory data. Session Overview.

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Module 5

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  1. Module 5 Laboratory Diagnostics, Specimen Collection, and Biosafety Issues

  2. Learning Objectives • Prepare and maintain collection kits • Collect and transport specimens safely and correctly • Manage laboratory specimens safely • Interpret laboratory data

  3. Session Overview • Specimen collection • Specimen storage, handling, shipping • Disposing infectious waste in the field • Safety precautions in the laboratory • Working with reference laboratories • Managing laboratory data

  4. The Specimen Collection Kit

  5. Specimen Collection Kit • Viral transport medium (VTM) • Preparing the specimen collection kit • Managing the specimen collection kit

  6. What is Viral Transport Medium? • Used in the collection of samples for viral isolation and testing • Prevents specimen from drying out • Prevents bacteria and fungi growth

  7. How to choose VTM • Can be made in a lab or purchased • Different types of VTM: • For collection of animal specimens • For viral isolation • For molecular testing • (Do not use phosphate-based media) • If VTM is not available, 100% ethanol can be used for molecular testing

  8. Storing VTM • Sterile collection vials containing 2-3 ml of VTM • Vials can be stored in a freezer at -20 ºC until use • Vials can be stored for short periods of time at 4 - 6 ºC

  9. Managing Media Stock • Keep records of when the VTM was made • Do not use vials if the liquid becomes cloudy

  10. Collection vials with VTM Polyester fiber-tipped applicators Sterile saline (0,85% NaCl) Sputum or mucus trap Tongue depressors Specimen collection cups or Petri dishes Transfer pipettes Secondary container Ice packs Items for blood collection Personal protective equipment Field collection forms A pen or marker for labeling samples Specimen Collection Kit

  11. Sputum Trap

  12. Polyester Fiber-Tipped Applicator • Should be drayon, rayon, or polyester-fiber swabs • Donotuse calcium alginated or cotton swabs nor ones with wooden sticks; they inhibit PCR

  13. Tongue Depressors

  14. 15 ml Conical Centrifuge Tubes

  15. Specimen Collection Cups and Petri Dishes collection cups petri dishes

  16. Transfer Pipettes

  17. Phlebotomy Supplies • Tourniquet • Disposable needles • Vacuum tubes with EDTA • Plastic needle holder • Alcohol and iodine swabs • Gauze • Band-aids • Biohazard sharps container

  18. Gloves Mask Gown Eye protection Personal Protective Equipment

  19. How to Manage Kits • Store specimen collection kits in a dry, cool place • Store specimen collection kit where it will be accessible after hours and on weekends

  20. How to Safely And Correctly Collect Samples

  21. Clinical Specimen Collection Responsibility • Usually a hospital staff function • May be a rapid responder function if hospital staff are unavailable • Designate at least one member of the team trained to collect specimens

  22. Clinical Specimen Sources Be prepared to collect specimens before you leave for the field • Suspected cases • Symptoms consistent with influenza • Contacts • Including people living or working with suspected cases

  23. What to Collect Preferred specimens • Nasal swabs • Throat swabs • When possible: Nasopharyngeal specimens Other specimens • Posterior pharyngeal swabs • Nasal washes • Acute and convalescent serum Collect the sample on several different days

  24. What to Collect From an Ambulatory patient • Nasal swab and • Throat swab • Can be collected into the same VTM From an Intubated patient • Lower respiratory aspirate

  25. When to Collect Respiratory Specimens • As soon as possible after symptoms begin • Before antiviral medications are administered • Even if symptoms began more than one week ago • Collect multiple specimens on multiple days

  26. Respiratory Specimens • Nasopharyngeal aspirate or swabs • Posterior pharyngeal swabs • Nasal swabs or nasal wash Nasopharyngeal aspirate and nasopharyngeal swabs have greatest sensitivity in detecting influenza virus

  27. Serological Samples Paired serum samples are most useful Acute sample Within 7 days after symptom onset Convalescent sample More than 21 days after symptom onset

  28. Alternative Diagnostic Testing If avian influenza is not the cause of the illness, consider the following alternative diagnostic testing: • Culturing for other respiratory viruses • SARS testing • Rapid influenza test • PPD for tuberculosis

  29. Personal Protective Equipment • Masks (N-95 or N/P/R-100) • Gloves • Protective eye ware (goggles) • Hair covers • Boot or shoe covers • Protective clothing (gown or apron)

  30. How to Collect Specimens

  31. Field Data Collection Form • Patient name • Unique identification number • Patient demographic information • Patient’s health status

  32. Specimen Tracking System Maintain a database to track: • Identification number • Subject information • Specimen collection date • Specimen collection location • Diagnostic test results

  33. Nasopharyngeal Swab • Insert dry swab into nostril and back to nasopharynx • Leave in place for a few seconds • Slowly remove swab while slightly rotating it

  34. Nasopharyngeal Swabcontinued • Use a different swab for the other nostril • Put tip of swab into vial containing VTM, breaking applicator’s stick

  35. Nasopharyngeal Swab Demonstration

  36. Posterior Pharyngeal Swab • Ask the subject to open his or her mouth • Depress the tongue • Swab the posterior pharynx • Avoid the tonsils

  37. Posterior Pharyngeal Swab Demonstration

  38. Nasopharyngeal AspirateCollection Process • Attach mucus trap to vacuum source • Place catheter into nostril parallel to palate • Apply vacuum • Slowly remove catheter while slightly rotating it • Repeat with other nostril using the same catheter • After collection, flush catheter with 3 ml VTM and return VTM to a plastic vial

  39. Nasopharyngeal Aspirate Demonstration

  40. How to Collect Blood • Put tourniquet on subject Photos: CDC/ Jim Gathany

  41. How to Collect Blood • Clean area with iodine and alcohol

  42. How to Collect Blood • Attach assemble needle, needle holder, and collection tube

  43. How to Collect Blood • Insert needle into vein Collect at least 2 ml blood

  44. How to Collect Blood • Remove needle and apply pressure

  45. How to Collect Blood • Dispose of needle in biohazard sharps container Do not reuse a needle

  46. Patient Care • After blood draw, apply light pressure • After removing gauze, place a sterile Band-Aid

  47. How to Label Samples Use pre-printed barcode labels: • On the specimen container • On the field data collection form • On the log book Label each specimen with: • Subject’s name • Subject’s unique identification number

  48. Personal Protective Equipment • Masks (N-95 or N/P/R-100) • Gloves • Protective eye ware (goggles) • Hair covers • Boot or shoe covers • Protective clothing (gown or apron)

  49. Antecubital Venipuncture(Blood Sample) Demonstration

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